2.3. Phytochemical Analysis
Qualitative phytochemical analyses of both the extracts were performed by following the protocol of Adetuyi and Popoola [26], Trease and Evans [27], and Sofowora [28].
Tannins. 200 mg of plant material was boiled in 10 mL distilled water and few drops of FeCl3 were added to the filtrate; a blue-black precipitate indicated the presence of Tannins.
Alkaloids. 200 mg plant material was boiled in 10 mL methanol and filtered. 1% HCl was added followed by 6 drops of Dragendorff reagent, and brownish-red precipitate was taken as evidence for the presence of alkaloids.
Saponins (Frothing test). 5 mL distilled water was added to 200 mg plant material. 0.5 mL filtrate was diluted to 5 mL with distilled water and shaken vigorously for 2 minutes. Formation of stable foam indicates the presence of saponins.
Cardiac Glycosides (Keller-Kiliani test). 2 mL filtrate was treated with 1 mL glacial acetic acid containing few drops of FeCl3.Conc. H2SO4 was added to the above mixture giving green-blue colour depicting the positive results for presence of cardiac glycosides.
Steroids (Liebermann-Burchard reaction). 200 mg plant material was added in 10 mL chloroform. Acetic anhydride was added in the ratio of 1 : 1 which resulted into the formation of blue-green ring pointing towards the presence of steroids.
Terpenoids (Salkowski test). To 200 mg plant material 2 mL of chloroform (CHCl3) and 3 mL of concentrated sulphuric acid (H2SO4) were carefully added. A reddish brown colouration signified the presence of terpenoids.
Flavonoids. To the aqueous filtrate 5 mL of dilute ammonia solution was added, followed by concentrated H2SO4. A yellow colouration indicated the presence of flavonoids.
Phlobatannins. The deposition of a red precipitate denoted the presence of phlobatannins when 200 mg of plant material was dissolved in 10 mL of aqueous extract and few drops of 1% HCl were added in the boiling tube.
Anthraquinones. 500 mg of dried plant leaves were boiled in 10% HCl for 5 mins and filtrate was allowed to cool. Equal volume of CHCl3 with few drops of 10% NH3 was added to 2 mL filtrate. The formation of rose-pink colour implies the presence of Anthraquinones.
Reducing Sugars. To the 10 mL of aqueous extract a few drops of Fehling’s solution A and B were added; an orange red precipitate suggests the presence of reducing sugars.